牙龈卟啉单胞菌脂多糖(Pg-LPS)对破骨细胞EphA2基因表达的调控

目的:研究小鼠单核巨噬细胞系RAW264.7在破骨分化过程中,Pg-LPS对破骨细胞EphA2表达的影响。方法:用终浓度10 mg/L的Pg-LPS 刺激RAW264.7 细胞后,分别在1、3、5 d,应用RT-PCR检测破骨细胞中EphA2基因和破骨细胞相关基因(破骨细胞内基质金属蛋白酶( MMP9)、ACP5、c-fos、组织蛋白酶K(CtsK)、NFATc1)的表达,并且通过酒石酸抗酸性磷酸酶(TRAP)染色观察实验组和对照组破骨细胞的分化成熟情况。结果:RT-PCR检测10 mg/L的Pg-LPS在第3天和5天,实验组比对照组EphA2基因表达分别增高2.4倍和1.2倍,两组之间存在显著差异(P<0.01);同时也能够促进破骨相关基因c-fos、NFATc1、CtsK、ACP5、MMP9的表达,实验组与对照组相比差异有显著性;TRAP染色结果显示:实验组比对照组的TRAP阳性多核细胞数目明显增多。结论:10 mg/L的Pg-LPS对小鼠单核巨噬细胞系RAW264.7,在破骨分化的中期和晚期均能够促进EphA2基因的表达,但是在破骨分化早期对EphA2基因的表达无明显作用。

Effect of Porphyromonas Gingivalis Lipopolysaccharide (Pg-LPS) on the Expression of EphA2 Gene in Osteoclast.

Objective: To investigate the effect of porphyromonas gingivalis lipopolysaccharide (Pg-LPS) on the expression of EphA2 gene in RAW264.7 cells during osteoclastic differentiation. Methods: RAW264.7 cells were stimulated with 10μg/ml of porphyromonas gingivalis lipopolysaccharide (Pg-LPS) at 1, 3 and 5 d. RT-PCR was applied to determine the expression of EphA2 gene and the osteoclast related genes(MMP9,c-fos,ACP5,CtsK,NFATc1). Tartrate-resistant acid phosphatase (TRAP) staining was applied to observe osteoclast differentiation and maturation. Results: Compared with the control group, at the 3 d and 5 d, the EphA2 gene mRNA expression was significantly increased 2.4-fold and 1.2-fold in Pg-LPS group. At the 1 d, there was no obvious difference between the Pg-LPS group and control group. Meanwhile Pg-LPS stimulation significantly promote osteoclast related genes of c-fos, NFATc1, CtsK, ACP5 and MMP9 expressions.Tartrate-resistant acid phosphatase (TRAP) staining showed that compared with the control group, the number of the TRAP positive cells was significantly increased in the Pg-LPS group. Conclusion: Pg-LPS can promote the expression of EphA2 gene in the middle and later stages of osteoclast differentiation,but has no obvious effect in the early stage.